B-Cell CD40 Expression by Flow Cytometry, Blood
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
Evaluating patients for hyper-IgM type 3 (HIGM3) syndrome due to defects in CD40, typically seen in patients <10 years of age
Assessing B-cell immune competence in other clinical contexts, including autoimmunity, malignancy and transplantation
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
CD40 by Flow, QL, B
Specimen Type Describes the specimen type needed for testing
Whole Blood EDTA
Specimen Required Defines the optimal specimen. This field describes the type of specimen required to perform the test and the preferred volume to complete testing. The volume allows automated processing, fastest throughput and, when indicated, repeat or reflex testing.
Specimen must arrive within 72 hours of draw. Send specimen Sunday through Thursday only. Draw and package specimen as close to shipping time as possible.
Container/Tube: Lavender top (EDTA)
Specimen Volume: 3 mL
Collection Instructions: Send specimen in original tube. Do not aliquot.
1. Ordering physician name and phone number are required.
2. For serial monitoring, we recommend that specimen draws be performed at the same time of day.
Specimen Minimum Volume Defines the amount of specimen required to perform an assay once, including instrument and container dead space. Submitting the minimum specimen volume makes it impossible to repeat the test or perform confirmatory or perform reflex testing. In some situations, a minimum specimen volume may result in a QNS (quantity not sufficient) result, requiring a second specimen to be collected.
Mild OK; Gross reject
Mild OK; Gross reject
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
|Whole Blood EDTA||Ambient||72 hours|
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
The adaptive immune response includes both cell-mediated (mediated by T cells and natural killer [NK] cells) and humoral (mediated by B cells) immunity. After antigen recognition and maturation in secondary lymphoid organs, some antigen-specific B cells terminally differentiate into antibody-secreting plasma cells. Decreased numbers or aberrant function of B cells result in humoral immune deficiency states with increased susceptibility to infections, and these may be either primary (genetic) or secondary immunodeficiencies. Secondary causes include medications, malignancies, infections, and autoimmune disorders (this does not cause immunodeficiency with increased infection).
CD40 is a member of the tumor necrosis factor receptor superfamily, expressed on a wide range of cell types including B cells, macrophages, and dendritic cells.(1) CD40 is the receptor for CD40 ligand (CD40LG), a molecule predominantly expressed by activated CD4+ T cells. CD40/CD40LG interaction is involved in the formation of memory B lymphocytes and promotes immunoglobulin (Ig) isotype switching.(1) CD40LG expression in T cells requires cellular activation, while CD40 is constitutively expressed on the surface of B cells and other antigen-presenting cells.
Hyperimmunoglobulin M (hyper-IgM or HIGM) syndrome is a rare primary immunodeficiency characterized by increased or normal levels of IgM with low IgG and/or IgA.(2) Patients with hyper-IgM syndromes may have genetic defects or mutations in 1 of 5 known genes. These genes are CD40LG, CD40, AICDA (activation-induced cytidine deaminase), UNG (uracil DNA glycosylase), and IKBKG (inhibitor of kappa light polypeptide gene enhancer in B cells, kinase gamma; also known as NEMO).(2) Not all cases of hyper-IgM syndrome fit into these known genetic defects. Mutations in CD40LG and IKBKG are inherited in an X-linked fashion, while mutations in the other 3 genes are autosomal recessive. Elevated IgM is only one of the features of NEMO deficiency and therefore, it is no longer classified exclusively with the hyper IgM syndromes.
Distinguishing between the different forms of hyper-IgM syndrome is very important because of differing prognoses. CD40 and CD40LG deficiency are among the more severe forms, which typically manifest in infancy or early childhood, and are characterized by an increased susceptibility to opportunistic pathogens (eg, Pneumocystis carinii, Cryptosporidium, and Toxoplasma gondii).(3)
CD40 deficiency, also known as hyper-IgM type 3 (HIGM3), accounts for <1% of hyper-IgM syndromes. Flow cytometry analysis shows complete lack of CD40 expression on the B cells of these patients.(4) Intravenous injection with IgG is the treatment of choice along with immune reconstitution with hematopoietic cell transplantation. To date, all documented CD40-deficient patients have been diagnosed before age 1. Consequently, when used in the context of HIGM3, this test is only indicated in children (for diagnosis). In the case of CD40L deficiency, this test can be used for male patients or in females of child-bearing age (to identify carriers). A larger age spectrum has been reported with CD40L deficiency, ranging from infancy to early adulthood.
CD40 expression on B cells is also an indicator of immune status (eg, after the use of biological immunomodulatory therapy for autoimmune disease, cancer and transplantation).
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
This assay is qualitative; CD40 expression is reported as present (normal) or absent (abnormal). Normal B cells express surface CD40 on the majority of cells.
Hyper IgM (HIGM3) syndrome patients typically do not express CD40 on the surface of B cells. Genotyping of CD40 is required for a definite diagnosis of HIGM3. Contact Mayo Medical Laboratories for ordering assistance.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
For questions about appropriate test selection, contact Mayo Medical Laboratories.
This test is not used to detect in CD40L expression (CD154), which is responsible for X-linked hyper IgM syndrome (HIGM1); see XHIM/82964 X-Linked Hyper IgM Syndrome, Blood.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Bishop GA, Hostager BS: The CD40-CD154 interaction in B cell-T cell liaisons. Cytokine Growth Factor Rev 2003;14:297-309
2. Lee WI, Torgerson TR, Schumacher MJ, et al: Molecular analysis of a large cohort of patients with hyper immunoglobulin M (IgM) syndrome. Blood 2005;105:1881-1890
3. Kutukculer N, Moratto D, Aydinok Y, et al: Disseminated cryptosporidium infection in an infant with hyper-IgM syndrome caused by CD40 deficiency. J Pediatr 2003;142:194-196
4. Ferrari S, Giliani S, Insalaco A, et al: Mutations of CD40 gene cause an autosomal recessive form of immunodeficiency with hyper IgM. Proc Natl Acad Sci USA 2001;98:12614-12619
Method Description Describes how the test is performed and provides a method-specific reference
The assay involves a multicolor panel of antibodies for the following markers: CD45, CD19, and CD40. After the staining, RBCs are lysed. The remaining cells are washed and analyzed by flow cytometry on a BD FACSCanto instrument. The cell surface expression of CD40 in B cells (CD19+) is determined and expressed as being present or absent. (Unpublished Mayo method)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday
Do not send specimen after Thursday. Specimen must be received by 10 a.m. on Friday.
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test
Test Classification Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer's instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR), Investigation Use Only (IUO) product, or a Research Use Only (RUO) product.
This test was developed using an analyte specific reagent. Its performance characteristics were determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.
CPT Code Information Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Medical Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.
LOINC® Code Information Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the result codes returned for this test or profile.
|Result ID||Reporting Name||LOINC Code|
|89009||CD40 by Flow, QL, B||In Process|